A proximity-labeling proteomic approach to investigate invadopodia molecular landscape in breast cancer cells

ABSTRACT: Invadopodia constitute a specialized machinery required for invasive tumor cells to travel through basement membranes and the interstitial matrix, and thus represent a key feature of metastatic cells. Therefore a better understanding of invadopodia biology and characteristics might permit the identification of markers of metastatic potential. Substantial efforts have been made to characterize invadopodia molecular composition, dynamics or triggering stimuli. However, the full molecular identity of these structures is still missing due to the fact that the isolation and purification of these structures has been challenging. To fill this gap, we developed a non-hypothesis driven proteomic approach based on the innovative BioID proximity biotinylation approach using the invadopodia-specific protein Tks5/FISH (its long form, Tks5) fused to the biotin ligase BirA as bait. As control, we also generated cells expressing the short form of Tks5 (Tks5) unable to localize to invadopodia. After validation of our cell models, Tks5 close neighbors were identified by label-free mass spectrometry after pulldown of biotinylated proteins. Known invadopodia components were identified revealing the pertinence of our strategy. Furthermore, this strategy allowed us to identify novel Tks5 in cellulo partners that appear as potential new invadopodia components/regulators and potential markers of metastasis.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Cell Culture

DISEASE(S): Breast Cancer

SUBMITTER: AUDEBERT Stephane  

LAB HEAD: Camoin Luc

PROVIDER: PXD015847 | Pride | 2020-04-24

REPOSITORIES: Pride